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Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene

Magnetic resonance (MR) reporter genes have the potential for tracking the biodistribution and fate of cells in vivo, thus allowing the safety, efficacy and mechanisms of action of cell‐based therapies to be comprehensively assessed. In this study, we evaluate the effectiveness of the iron importer...

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Autores principales: Pereira, Sofia M., Herrmann, Anne, Moss, Diana, Poptani, Harish, Williams, Steve R., Murray, Patricia, Taylor, Arthur
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4981909/
https://www.ncbi.nlm.nih.gov/pubmed/26929139
http://dx.doi.org/10.1002/cmmi.1686
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author Pereira, Sofia M.
Herrmann, Anne
Moss, Diana
Poptani, Harish
Williams, Steve R.
Murray, Patricia
Taylor, Arthur
author_facet Pereira, Sofia M.
Herrmann, Anne
Moss, Diana
Poptani, Harish
Williams, Steve R.
Murray, Patricia
Taylor, Arthur
author_sort Pereira, Sofia M.
collection PubMed
description Magnetic resonance (MR) reporter genes have the potential for tracking the biodistribution and fate of cells in vivo, thus allowing the safety, efficacy and mechanisms of action of cell‐based therapies to be comprehensively assessed. In this study, we evaluate the effectiveness of the iron importer transferrin receptor‐1 (TfR1) as an MR reporter gene in the model cell line CHO‐K1. Overexpression of the TfR1 transgene led to a reduction in the levels of endogenous TfR1 mRNA, but to a 60‐fold increase in total TfR1 protein levels. Although the mRNA levels of ferritin heavy chain‐1 (Fth1) did not change, Fth1 protein levels increased 13‐fold. The concentration of intracellular iron increased significantly, even when cells were cultured in medium that was not supplemented with iron and the amount of iron in the extracellular environment was thus at physiological levels. However, we found that, by supplementing the cell culture medium with ferric citrate, a comparable degree of iron uptake and MR contrast could be achieved in control cells that did not express the TfR1 transgene. Sufficient MR contrast to enable the cells to be detected in vivo following their administration into the midbrain of chick embryos was obtained irrespective of the reporter gene. We conclude that TfR1 is not an effective reporter and that, to track the biodistribution of cells with MR imaging in the short term, it is sufficient to simply culture cells in the presence of ferric citrate. Copyright © 2016 The Authors Contrast Media & Molecular Imaging Published by John Wiley & Sons Ltd.
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spelling pubmed-49819092016-08-26 Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene Pereira, Sofia M. Herrmann, Anne Moss, Diana Poptani, Harish Williams, Steve R. Murray, Patricia Taylor, Arthur Contrast Media Mol Imaging Full Papers Magnetic resonance (MR) reporter genes have the potential for tracking the biodistribution and fate of cells in vivo, thus allowing the safety, efficacy and mechanisms of action of cell‐based therapies to be comprehensively assessed. In this study, we evaluate the effectiveness of the iron importer transferrin receptor‐1 (TfR1) as an MR reporter gene in the model cell line CHO‐K1. Overexpression of the TfR1 transgene led to a reduction in the levels of endogenous TfR1 mRNA, but to a 60‐fold increase in total TfR1 protein levels. Although the mRNA levels of ferritin heavy chain‐1 (Fth1) did not change, Fth1 protein levels increased 13‐fold. The concentration of intracellular iron increased significantly, even when cells were cultured in medium that was not supplemented with iron and the amount of iron in the extracellular environment was thus at physiological levels. However, we found that, by supplementing the cell culture medium with ferric citrate, a comparable degree of iron uptake and MR contrast could be achieved in control cells that did not express the TfR1 transgene. Sufficient MR contrast to enable the cells to be detected in vivo following their administration into the midbrain of chick embryos was obtained irrespective of the reporter gene. We conclude that TfR1 is not an effective reporter and that, to track the biodistribution of cells with MR imaging in the short term, it is sufficient to simply culture cells in the presence of ferric citrate. Copyright © 2016 The Authors Contrast Media & Molecular Imaging Published by John Wiley & Sons Ltd. John Wiley and Sons Inc. 2016-02-29 2016 /pmc/articles/PMC4981909/ /pubmed/26929139 http://dx.doi.org/10.1002/cmmi.1686 Text en Copyright © 2016 The Authors. Contrast Media & Molecular Imaging published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Full Papers
Pereira, Sofia M.
Herrmann, Anne
Moss, Diana
Poptani, Harish
Williams, Steve R.
Murray, Patricia
Taylor, Arthur
Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title_full Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title_fullStr Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title_full_unstemmed Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title_short Evaluating the effectiveness of transferrin receptor‐1 (TfR1) as a magnetic resonance reporter gene
title_sort evaluating the effectiveness of transferrin receptor‐1 (tfr1) as a magnetic resonance reporter gene
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4981909/
https://www.ncbi.nlm.nih.gov/pubmed/26929139
http://dx.doi.org/10.1002/cmmi.1686
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